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Chimonanthus of Calycanthaceae is a small endemic genus in China, with unusual winter-blooming sweet flowers widely cultivated for ornamentals and medicinal uses. The evolution of Chimonanthus plastomes and its phylogenetic relationships remain unresolved due to limited availability of genetic resources. Here, we report fully assembled and annotated chloroplast genomes of five Chimonanthus species. The chloroplast genomes of the genus (size range 153,010 – 153,299 bp) reveal high similarities in gene content, gene order, GC content, codon usage, amino acid frequency, simple sequence repeats, oligonucleotide repeats, synonymous and non-synonymous substitutions, and transition and transversion substitutions. Signatures of positive selection are detected in atpF and rpoB genes in C. campanulatus. The correlations among substitutions, InDels, and oligonucleotide repeats reveal weak to strong correlations in distantly related species at the intergeneric levels, and very weak to weak correlations among closely related Chimonanthus species. Chloroplast genomes are used to reconstruct a well-resolved phylogenetic tree, which supports the monophyly of Chimonanthus. Within Chimonanthus, C. praecox and C. campanulatus form one clade, while C. grammatus, C. salicifolius, C. zhejiangensis, and C. nitens constitute another clade. Chimonanthus nitens appears paraphyletic and is closely related to C. salicifolius and C. zhejiangensis, suggesting the need to reevaluate the species delimitation of C. nitens. Chimonanthus and Calycanthus diverged in mid-Oligocene; the radiation of extant Chimonanthus species was dated to the mid-Miocene, while C. grammatus diverged from other Chimonanthus species in the late Miocene. C. salicifolius, C. nitens(a), and C. zhejiangensis are inferred to have diverged in the Pleistocene of the Quaternary period, suggesting recent speciation of a relict lineage in the subtropical forest regions in eastern China. This study provides important insights into the chloroplast genome features and evolutionary history of Chimonanthus and family Calycanthaceae.

In order to better understand the floral fragrance compounds of Chimonanthus praecox belonging to genus Chimonanthus of Chimonanaceae in Yunnan, headspace solid-phase microextraction combined with gas chromatography-mass spectrometry was used to analyze these compounds from four C. praecox plants with different floral colors. Thirty-one types of floral fragrance compounds were identified, among which terpenes, alcohols, esters, phenols, and heterocyclic compounds were the main compounds. Interestingly, the floral fragrance compounds identified in the flowers of C. praecox var. concolor included benzyl acetate, α-ocimene, eugenol, indole, and benzyl alcohol. By contrast, the floral fragrance compounds β-ocimene, α-ocimene, and trans-β-ocimene were detected in C. praecox var. patens. Cluster analysis showed that C. praecox var. concolor H1, H2, and C. praecox var. patens H4 were clustered in one group, but C. praecox var. patens H3 was individually clustered in the other group. Additionally, principal component analysis showed that α-ocimene, benzyl alcohol, benzyl acetate, cinnamyl acetate, eugenol, and indole were the main floral fragrance compounds that could distinguish the four C. praecox with different floral colors in Yunnan. This study provides a theoretical basis for further elucidating the mechanism and pathway of the floral fragrance release of C. praecox.

Background Three species of wintersweets: Chimonanthus salicifolius S. Y. Hu, Chimonanthus zhejiangensis M. C. Liu and Chimonanthus grammalus M. C. Liu are widely distributed in China. The three wintersweets belonging to the genus of Chimonanthus that can synthesize abundant terpenoids that are beneficial to human health. Their buds and leaves are traditional Chinese herb applied by the ‘She’ ethnic minority in southeast of China. Squalene is a multi-functional and ubiquitous triterpene in plants, which is biosynthesized by squalene synthase (SQS) using farnesyl diphosphate (FPP) as the substrate. The synthesis of squalene in wintersweet was not clearly. This work would provide us much help to further understand the terpene metabolism in wintersweet and its health function to people at phytochemistry and molecular levels. Results In this study, we identified squalene component in the extractions of leaves of three wintersweets and isolated SQS genes from leaf transcriptomes. The three SQSs were highly conservative, so CzSQS from C. zhejiangensis was just determined the enzymatic activity. The in vitro expressed CzSQS that deleted two transmembrane domains could catalyze FPP to generate squalene with the presence of NADPH and Mg2+. Conclusions The squalene was one of wintersweet leaves phytochemicals. The squalene synthases of three wintersweet plants were highly conserved. The CzSQS was capable to catalyze two FPP molecules to squalene. Electronic supplementary material The online version of this article (10.1186/s40529-018-0246-6) contains supplementary material, which is available to authorized users.

Chimonanthus Lindl. has seven family members with Ch. praecox blooming in winter and others in autumn. The plants are widely used in landscaping, fragrance industry, as well as production of medicinal material because of their flower fragrance. Both Chimonanthus flowers and leaves contain abundant secondary metabolites. But it is unclear whether the flowers from different species have distinct ingredients and antioxidant activity. To address the question, we evaluated the main compositions of the flowers from five Chimonanthus species and analyzed the antioxidant activity of the compositions. The results indicated that flavonoids and coumarins are the two major types of substances with antioxidant capacity in Chimonanthus flowers. Totally nine antioxidative compounds, including scopolin and scopoletin, were identified based on the high performance liquid chromatography-photo diode array detection-electrospray ionization tandem mass spectrometry (HPLC-DAD/ESI–MS). All the antioxidant compounds were confirmed by pre-column 2, 2-diphenyl-1- picryhydrazyl (DPPH) assay. Rutin (highest in content), isoquercitrin and quercetin appeared stronger scavenging DPPH radical ability than the other five compounds, mainly due to their ortho-dihydroxyl groups. Among the five Chimonanthus species, Ch. praecox flower is most abundant in secondary metabolites and shows most potent antioxidant activity. The other four flowers contain the same types of ingredients with Ch. praecox, but exhibit lower antioxidant activity. The results obtained from this comprehensive analysis provide a theoretical basis for the utilization of the plant resources of Chimonanthus genus.

Chimonanthus nitensOliv. (CNO), having been studied and developed as the tea beverages, tea raw materials and preparations, an unique species in China, have been extensively used for treating colds and influenza for centuries. In the present study, a method based on the modified mass defect filter (MDF) was firstly developed and validated for comprehensive profiling of coumarins in the different parts of CNO via ultra-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS). The five-point screening approach based on MDF and the visual isotopic ion technique could rapidly screen the interested precursor ions. The fragmentation behavior of coumarins was systematically investigated and a total of 42 coumarins including 27 potential new ones were unambiguously or tentatively identified in the CNO. Collectively, the results obtained in the present work may provide useful information for future utilization of CNO.

Chimonanthus campanulatus R.H. Chang & C.S. Ding is a good horticultural tree because of its beautiful yellow flowers and evergreen leaves. In this study, fluorescence in situ hybridization (FISH) was used to analyse mitotic metaphase chromosomes of Ch. campanulatus with 5S rDNA and (AG3T3)3 oligonucleotides. Twenty-two small chromosomes were observed. Weak 5S rDNA signals were observed only in proximal regions of two chromosomes, which were adjacent to the (AG3T3)3 proximal signals. Weak (AG3T3)3 signals were observed on both chromosome ends, which enabled accurate chromosome counts. A pair of satellite bodies was observed. (AG3T3)3 signals displayed quite high diversity, changing in intensity from weak to very strong as follows: far away from the chromosome ends (satellites), ends, subtelomeric regions, and proximal regions. Ten high-quality spreads revealed metaphase dynamics from the beginning to the end and the transition to anaphase. Chromosomes gradually grew larger and thicker into linked chromatids, which grew more significantly in width than in length. Based on the combination of 5S rDNA and (AG3T3)3 signal patterns, ten chromosomes were exclusively distinguished, and the remaining twelve chromosomes were divided into two distinct groups. Our physical map, which can reproduce dynamic metaphase progression and distinguish chromosomes, will powerfully guide cytogenetic research on Chimonanthus and other trees.

Voltammetric scan can record the profile of electrochemical active substances in plant tissues. Because the distribution of chemical components in plants is controlled by genes, these profiles can reflect differences at the genetic level in different species. In this study, the voltammetric scan was applied to the investigation of macrophanerophytes taxonomy. All species of Chimonanthus with two exotaxa were deliberately selected due to their controversial infrageneric relationship. Electrode surface modification was excluded in this work to improve the convenience and accuracy of the fingerprint recording process. The dendrogram deduced from the electrochemical fingerprint data suggests that Ch. Zhejiangensis and Ch. grammatus are two groups of Ch. nitens, which may be only the ecotype of Ch. nitens, rather than independent taxonomic species. The small variations between the three species may be due to environmental factors and cannot be used for species formation. In addition, Ch. campanulatus and Ch. Praecox were clustered together with a close relationship.

Chimonanthus plants widely distributed in southern area of China, which have a long history of edibles and medicine. Phytochemical investigations have shown that Chimonanthus produced 143 non-volatile constituents, including alkaloids, flavonoids, terpenoids, coumarins and others, which exhibit significant anti-oxidant, anti-bacterial, anti-cancer, anti-inflammatory, antihyperglycemic, antihyperlipidemic and other biological activities. On the basis of systematic reviewing of literatures, this article overviews the non-volatile constituents and pharmacology of Chimonanthus from domestic and foreign over the last 30 years (until June 2018), and may provide a useful reference for the further development of Chimonanthus.

Chimonanthus grammatus M.C.Liu is an endangered and national-protected medicinal plant species with highly restricted and disjunctive geographical distribution in the border area of Anyuan and Huichang counties of Jiangxi province, China. In this study, six natural populations of the C. grammatus (AY1, AY2, and AY3 from Anyuan; HC1, HC2, and HC3 from Huichang county) and one cultivated population of Chimonanthus praecox (as one outgroup) from Nanchang (NC) city, Jiangxi, were sampled to analyze its genetic diversity and structure using ten microsatellite (SSR) markers. Results showed that Nei's genetic diversity values (He) of seven populations ranged from 0.574 to 0.742, with an average of 0.666 ± 0.028. Values of Shannon's information index ( I ) per population ranged from 1.215 to 1.832, with an average of 1.535 ± 0.074. For C. grammatus species, AY1 population from Anyuan county harbored the highest genetic diversity ( I = 1.832 ± 0.195, He = 0.742 ± 0.061), while HC3 population from Huichang county had the lowest levels of genetic diversity ( I = 1.358 ± 0.208, He = 0.601 ± 0.078), respectively. The low genetic differentiation (Fst = 0.089) was detected in C. grammatus species. UPGMA and PCA analyses classified the seven populations into three branches: AY1, AY2 and AY3 were clustered together; HC1, HC2, and HC3 were grouped together; and NC was clustered by itself. These results revealed that ten SSR markers were effective to investigate the genetic diversity of C. grammatus and to genetically differentiate it from another species in Chimonanthus .

To investigate interspecific cross-compatibility among Chimonanthus praecox (L.) Link, C. salicifolius S. Y. Hu, and C. nitens Oliver, pollen–pistil interaction by fluorescence microscopy and hybrid embryo development by whole mount clearing technique were observed. The results indicated that for all reciprocal crosses among the three species, pollen grains of the male parents could adhere to and germinate in the stigmas of female parents. For any cross combination, pollen tube growth could reach the embryo sac, and double fertilization could be completed. These findings indicated that no obvious barriers to interspecific hybridization existed at the stages of pollen germination and pollen tube elongation until double fertilization. Although certain proportion (29.6% at 9 d and 36.3% at 15 d after pollination) of abortions (e.g., non-fertilization and retarded or non-existent endosperm development) in normally pollinated embryos of C. praecox at different stages were also observed, a greatly increased proportion (66.1% at 8 d and 76.7% at 15 d after pollination in C. praecox × C. nitens, and 60.0% at 8 d and 75.0% at 15 d after pollination in C. praecox × C. salicifolius) of hybrid embryos did not complete double fertilization or completed double fertilization but underwent abortion due to the retardation of endosperm development. Comparing to normally pollinated embryos of C. praecox, the retardation of endosperm development in hybridizations occurred earlier. Moreover, the occurrence of abortions among different cross combinations was basically consistent. Thus, the retardation of the endosperm development of hybrid embryos after double fertilization (after the globular embryo stage) is an important factor leading to the failure of distant hybridization between Chimonanthus species.

The aim of this study was to investigate the interventional effect of the Chimonanthus salicifolicus essential oil on murine chronic stress-induced anxiety. Rats model of uncertainty empty bottle was established to induce anxiety. The exploratory change, grooming and attack behaviors were observed. After 7 days training period followed by continuous 14 days’ stimulation, rats serum level of glucose (GLU), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were detected. The results showed that rat model of uncertainty empty bottle induced anxiety well. C. salicifolicus essential oil smelling could significantly appease the anxiety of rat which include the reduction of attacking behaviors. It also showed a reduced effect to the level of TC and GLU. The result also indicated that C. salicifolicus essential oil might have an anti-anxiety effect on rat uncertainty empty bottle model and could be used as a potential. Key words: Anxiolytics, Chimonanthus salicifolicus essential oil, chronic emotional stress.

Sinocalycanthus chinensis, an endangered species endemic to China, is cultivated as an ornamental landscape tree in China. However, S. chinensis, Chimonanthus species and Calycanthus floridus are difficult to be distinguished in seedling market because of their similar morphological characters. In this study, ISSR (inter-simple sequence repeats) were applied to detect S. chinensis from its closely related species. A unique 748-bp band was found in all accessions of S. chinensis. SCAR (sequence characterized amplified regions) markers were created by cloning and sequencing the specific band, and designing a pair of primers to amplify the band of 748 bp. Diagnostic PCRs were performed using the primer pair with the total DNAs of S. chinensis, Chimonanthus species and C. floridus as templates, with only S. chinensis being able to be amplified. This amplification is not only rapid (results can be obtained in less than 3 h), but is also easy to perform. Hence it is a feasible method for identifying S. chinensis in seedling market.

Based on nuclear and chloroplast sequences we resolve species relationships in Calycanthaceae and develop a biogeographic hypothesis that explains their intercontinental disjunctions and intra-continental diversification in eastern Asia. Fossil-calibrated penalized likelihood and Bayesian divergence time estimates indicate that the Northern Hemisphere Calycanthus and Chimonanthus diverged from each other in the mid-Miocene, while the Australian Idiospermum had already diverged by the Upper Cretaceous and likely represents a remnant of a former Gondwanan distribution of Calycanthaceae that included South America, as indicated by the occurrence of Cretaceous Calycanthaceae fossils in Brazil. Relationships within Calycanthus were difficult to resolve, but a shared 155-bp deletion in the trnL-F intergenic spacer unites the two North American species, which were also sisters in a cpDNA restriction site study. Their ancestor apparently crossed the Bering land bridge in the Miocene. The six species of Chimonanthus, by contrast, diverged from each other as recently as 1-2my ago, and a DIVA analysis with four areas of endemism recognized within China suggests three vicariance and two dispersal events within Chimonanthus, with initial vicariance having occurred between eastern and southwestern or central China. Further divergence then appears to have involved eastern and south-central China, and southwestern and central China.

Chimonanthus grammatus M.C.Liu is an endangered and national-protected medicinal plant species with highly restricted geographical distribution and disjunctly locates in the border area of Anyuan and Huichang county of Jiangxi province, China. In this study, we developed ten microsatellite loci for this species for the first time. Nine of them was screened by combining biotin capture method to enrich (CA)15 /(GA)15 microsatellites, and one primer is designed from one EST sequence (DW222670) of Chimonanthus praecox. Ten microsatellite primers were found to be polymorphic through screening 30 individuals. Na was from two to eighteen. Ho and He ranged from 0.233 to 0.933 and 0.326 to 0.898, respectly. Polymorphic information content (PIC) was from 0.316 to 0.890. Ten microsatellite primers will be useful to investigate the genetic structure of Chimonanthus grammatus M.C.Liu and to explain its morphological differentiation from another species in Chimonanthus.

Chimonanthus grammatus is an endangered and national-protected medicinal plant species with highly restricted geographical distribution and disjunctly locates in the border area of Anyuan and Huichang county of Jiangxi province, China. In this study, inter-simple sequence repeats (ISSR) markers were used to analyzed genetic diversity of seven populations of C. grammatus, collected from the area of Anyuan (4 populations) and Huichang (3 population) county of Jiangxi province, China. Our results showed that ten primers produced 74 discernible bands (39 polymorphic), with a low genetic diversity at species level percentages of polymorphic bands (PPB = 52.7%, H = 0.157, I = 0.241) and at population level (PPB = 35.7%,H = 0.119, I = 0.177). A high population differentiation (Gst = 0.249) was detected among seven populations and significant association were found between genetic and geographical distances (r = 0.712, p < 0.001), suggesting that gene flow was restricted geographically. Analysis of molecular variance (AMOVA) also revealed that 26.4% of the ISSR variation resided among populations, while 73.6% resided within populations, and 21.8% of the ISSR variation resided among Anyuan and Huichang groups, while 78.2% resided within groups. Factors, such as gene flow, genetic drift and evolutionary history, might have important influence on genetic structure and diversity of C. grammatus populations. An understanding of the genetic diversity and population structure of C. grammatus can provide insight into the conservation and management of this species. Key words: Chimonanthus grammatus, genetic diversity, inter-simple sequence repeats (ISSR), conservation.

OBJECTIVE: Species containing extremely aromatic compounds in leaves of Chimonanthus was analyzed to evaluate its genetic diversity and genetic relationships. METHOD: AFLP molecular marker technique was used in the study, UPGMA cluster analysis was conducted with the software of POPGENE32. RESULTS: Five hundred and fifty-nine bands were amplified by 10 pairs of primers screened, of which 226 bands were polymorphic, and the percentage of polymorphic bands was 36.8%. Observed number of alleles, effective number of alleles, Nei's genetic diversity index and Shannon's information index were 1.992 6, 1.306 5, 0.199 2 and 0.325 1, respectively. Genetic distances of the 21 populations were ranged from 0.039 2 to 0.289 4. CONCLUSION: Species containing extremely aromatic compounds in leaves of Chimonanthus with low genetic diversity play an important role in enhancing the protection of species and germplasm resources. Form the molecular level, the studies demonstrated the correctness of the result by Zhang Ruohui that species containing extremely aromatic compounds in leaves of Chimonanthus were divided into Ch. salicifolius, Ch. Zhejiangensis, Ch. nitens and Ch. grammatus.

A morphological comparison of Calycanthus L., Chimonanthus Lindl., and Sinocalycanthus W.C. Cheng and S.Y. Chang is presented. Intergeneric crosses between Sinocalycanthus chinensis (W.C. Cheng and S.Y. Chang) W.C. Cheng and S.Y. Chang and Calycanthus floridus L. produced hybrid progeny. Data are presented comparing the parents and the progeny. ×Sinocalycalycanthus raulstonii Lasseigne and Fantz is proposed as the name for these hybrids and is validated with a Latin diagnosis. The original cultivar is named `Hartlage Wine'.

Calycanthus praecox Linnaeus, Species Plantarum, ed. 2, 1: 718. 1762. "Habitat in Japonia." RCN: 3813. Holotype (Nicely in Castanea 30: 74. 1965): [icon] "Obai seu Robai" in Kaempfer, Amoen. Exot. Fasc.: 878, 879. 1712 (see p. 19). Current name: Chimonanthus praecox ( L.) Link (Calycanthaceae)., Published as part of Jarvis, Charlie, 2007, Chapter 7: Linnaean Plant Names and their Types (part C), pp. 370-473 in Order out of Chaos. Linnaean Plant Types and their Types, London :Linnaean Society of London in association with the Natural History Museum on page 376, DOI: 10.5281/zenodo.291971

A new pyrrolidinoindoline-type alkaloid, CPC-1, and a new tryptamine-derived dimeric alkaloid, CPC-2, were isolated from the seeds and rinds of Chimonanthus praecox f. concolor. The structure including the absolute configuration of CPC-1 was determined by chiral total synthesis. CPC-2 has a unique hexahydropyrroloquinoline skeleton.

Two new tryptamine-related alkaloids, chimonamidine and chimonanthidine, were isolated from the seeds of Chimonanthus praecox Link. and their structures including absolute configuration were elucidated by spectroscopic analysis and biomimetic total synthesis from tryptamine.

U razdoblju od 1998–2000. godine u Botaničkome vrtu Prirodoslovno-matematičkoga fakulteta Sveučilišta u Zagrebu bilježeni su vrijeme početka cvjetanja, vrhunac i kraj cvjetanja nekih vrsta koje cvjetaju tijekom zime ili u rano proljeće. Riječ je o vrstama iz rodova Chimonanthus, Cornus, Corylopsis, Forsythia, Hamamelis, Jasminum, Lonicera i Sarcococca. Pri tome je ustanovljeno da se za vrste Chimonanthus praecox, Jasminum nudiflorum, Lonicera fragrantissima i Lonicera purpusii, koje u povoljnim vremenskim uvjetima cvjetaju tijekom cijele zime, ne može unaprijed predvidjeti vrijeme početka, a često ni vrhunca cvjetanja. Vrijeme cvjetanja je u ovih vrsta pod vrlo snažnim utjecajem vremenskih prilika, ponajprije srednjih dnevnih i srednjih mjesečnih vrijednosti temperature zraka. Druge vrste (Sarcococca hookeriana var. humilis, S. saligna, Corylopsis sinensis var. glandulifera, većina vrsta roda Forsythia, Lonicera praeflorens) cvjetaju svake godine približno u isto vrijeme i na njih djeluju samo veće klimatske promjene, posebno više temperature u drugom dijelu zime. Njima su bliske vrste treće skupine (Cornus mas, Corylopsis spicata, C. willmottiae, Hamamelis japonica 'Zuccariniana', Forsythia viridissima) koje su tijekom tri godine cvjetale praktično u isto vrijeme. Općenito možemo zaključiti da vrste koje cvjetaju ranije pokazuju veću varijabilnost u pogledu vremena cvjetanja nego one koje cvjetaju kasnije (krajem zime ili početkom proljeća). Pri tome sve vrste u pravilu pokazuju najveću stabilnost u pogledu vremena završetka cvjetanja, nešto manje vrhunca cvjetanja, dok je vrijeme početka cvjetanja najvarijabilnije.

A B S T R A C T The floral anatomy of Idiospermum australiense (Diels) S. T. Blake is described from studies on relatively mature but unopened flower buds. The vascular system and general morphology of the flower is compared with that of Calycanthus and Chimonanthus (Calycanthaceae), especially with Calycanthus, the genus in which Diels originally placed Idiospermum. Inverted cortical bundles are present in both taxa but occur in different patterns. The tepals of Calycanthus are all 3-trace, whereas in Idiospermum only the upper tepals have three traces, the lower tepals having five to seven. All tepals of Calycanthus are spirally arranged; the lower tepals of Idiospermum are opposite or decussate in 1-3 pairs. The axial and recurrent bundle systems in Calycanthus are discrete except where the axial system turns downward, but in Idiospermum these systems, together with cortical bundles, are largely intermixed. The vascular supply to the carpel of Idiospermum, while possibly a modification of that of the carpels of Calycanthus, could also be interpreted as having an independent origin. Other differences and resemblances are described.

Vegetative structural features of Idiospermum, a monotypic ranalean genus rediscovered in 1972, are described and contrasted with comparable features of Calycanthus and Chimonanthus (Calycanthaceae) to note resemblances and differences. The leaf trace pattern and nodal anatomy of Calycanthaceae and Idiospermaceae (one-trace and unilacunar) are very similar. The nodal anatomy of certain highly reduced leaves (scale leaves) of Idiospermum is, however, two-trace and unilacunar. The microscopic wood anatomy of Idiospermum is distinctively different in a number of respects from that of the Calycanthaceae. Although there are similarities, it is suggested that the floral anatomy and vegetative features of Idiospermum set it apart sufficiently from the Calycanthaceae to justify family rank as the Idiospermaceae. THE REDISCOVERY of a new monotypic genus of dicotyledons, named Idiospermum australiense by Blake (1972), is of considerable interest. This rain forest tree of northeastern Australia was assigned by Blake to a new monotypic family, Idiospermaceae. It is an evergreen tree growing to 40 m high and 90 cm in diam. Its simple leathery leaves are petiolate, opposite, decussate (1/2 phyllotaxy), 12-26 cm long, 8-12 cm wide, oblong to elliptic and shortly acuminate with entire margins. The genus is of taxonomic importance because it is a new member of what is frequently referred to as the ranalean complex, essentially equivalent to the subclass Magnoliidae of Cronquist (1968) or to the superorder Annoniflorae of Thorne (1968, 1976). These taxa embrace a group of families viewed as having retained certain features of ancient and possibly ancestral flowering plants. In a previous paper (Wilson, 1976) I reported on the floral anatomy of Idiospermum, comparing it with that of Calycanthusfloridus L. and Chimonanthus praecox (L.) Link. In what follows, the results of certain studies on the vegetative morphology of this plant are presented, again comparing certain features with those of the same species of Calycanthus and Chimonanthus. The features investigated include the scale I Received for publication 14 June 1978; revision accepted 13 October 1978. I am indebted to Mr. B. P. Hyland, Commonwealth Scientific and Industrial Research Organization (Australia), for wood of Idiospermum; to Mr. R. W. Johnson, Queensland Herbarium, for seeds of Idiospermum, and to Dr. J. M. Fogg, Arboretum Barnes Foundation, for material of Chimonanthus. I am grateful to Prof. William T. Jackson, Dartmouth, for the photomicrographs, to Prof. J. P. Poole, Dartmouth, for reading the manuscript, and to Prof. A. E. DeMaggio, Dartmouth, for reading the manuscript and for other assistance. leaves of Idiospermum, the leaf trace pattern in Calycanthus and Idiospermum, and the nodal anatomy and wood of Calycanthus, Chimonanthus, and Idiospermum. MATERIALS AND METHODs-Material of Calycanthus was obtained from greenhouse-grown seedlings and mature plants in cultivation out-ofdoors (Calycanthus floridus L. var. floridus; voucher Wilson 880, in Jesup Herbarium, Dartmouth College), material of Chimonanthus from plants in cultivation, and material of Idiospermum from plants grown in the greenhouse. Wood samples of Idiospermum came from a tree in the native habitat. Procedures included the use of NaOH and chloral hydrate for clearing, especially in the study of scale leaves and nodes. In the study of the leaf trace patterns, terminal parts of shoots were taken, including 5-7 nodes below the apex, but omitting several of the younger uppermost nodes. This involved distances up to 18 cm. The stems were cut into pieces approximately 1.5 cm long, embedded in paraffin, sectioned at 12 ,tm in their proper order, stained with safranin and fast green, and permanently mounted. Three shoots of both Calycanthus and Idiospermum were prepared for study in this way. Following the leaf trace pattern even in mature shoots was made possible by prolonged soaking of the embedded pieces, one end exposed, in a mixture of glycerine and a detergent. A transection of the internodal region of Calycanthus and Idiospermum near the apex of a shoot shows that the vascular cylinder is organized into what has been termed a pseudosiphonostele (Bailey and Nast, 1948). This appears as an essentially continuous ring of primary phloem and primary xylem (Fig. 6, 7) interrupted

Publisher Summary This chapter describes the alkaloids of calycanthaceae. The family Calycanthaceae is represented by only two genera, Calycanthus L., with about four species in the United States, and Chimonanthus Lindl. ( Meratia Loisel. ) with two Asiatic species. Its relation to other plant families is uncertain, and the nature of its chemical constituents has not shed any light on possible affinities. The belief in the presence of an indole nucleus in calycanthine was so firmly rooted that the first structures that bear some resemblance to the true one still retained the indole nucleus, even though they are largely based on the then-known structure of calycanine. There are few examples in the history of organic chemistry where deductions of structure based on identified fragments have been as wide of the mark as in the work on calycanthine. Reactions, that because of their mildness could be presumed to proceed without molecular rearrangements, yielded products that only served to confuse the real structure.

Publisher Summary Indole has been isolated from the flowers of many Jasminium and Citrus species, from Robinia pseudacacia L., Cheirunthus cheiri L., Narcissus jonquilla L., and Chimonanthus fragrans Lindl.; it appears to be an essential constituent of the perfumes of these flowers. Reports have also been made of the isolation of indole from nonfloral material, for example, Celtis reticulosa Miq. and Thlaspi arvense L., but these have been criticized on the grounds that the indole was probably the result of bacterial action on a labile indole precursor. Numerous other claims for the presence of indole in plants have been based solely on the color reactions given either by the plant extracts or by the volatile constituents of the oils obtained from the blossoms by enfleurage. In some plants, at least, the reverse may be true, and indole may be converted into tryptophan by combination with serine. Whether this is the principal mode of biosynthesis of tryptophan remains to be determined; it is perhaps more likely that indole and tryptophan are products of alternative pathways of metabolism of indole-3-glycerol phosphate, and that conversion of the last-named into tryptophan does not proceed by way of indole.

Leaf flavonoids of Idiospermum australiense were examined and com- pared with those of Calycanthus and Chimonanthus (Calycanthaceae). Flavonoid data, in conjunction with morphological and anatomical features, were used to elucidate the phylogenetic relationships of the taxa. Five flavonol glycosides, based upon the aglycones quercetin and kaempferol, were isolated and identified from Calycanthus occidentalis, C. fioridus, and Chimonanthus praecox. Two flavones (7-0-diglucosides of apigenin and luteolin) and a di-C-glycosylflavone were detected in Idiospermum. No intermediate flavonoid patterns between the two groups were found. We feel that the distinctive nature of the flavonoids of Idiospermum, together with the morpho- logical and anatomical differences between the Calycanthaceae and Idiospermum that have been reported, support removing Idiospermum from the Calycanthaceae and placing it in a separate family, the Idiospermaceae.

Conversion of (±)-[β-14C, 2-3H]tryptophan and [β-14C, 2-3H]tryptamine into (–)-chimonanthine has been shown to occur in Chimonanthus fragrans(praecox) without loss of tritium. Degradation of the alkaloid has established the predicted labelling pattern. A synthesis of [2-3H]- and [2-3H]-indole from indoxyl is recorded and the structure of diacetoxymercuri-indole is revised.